Identification and Validation of Four Novel Promoters for Gene Engineering with Broad Suitability across Species

( Cai-yun Wang ),( Li-cheng Liu ),( Ying-cai Wu ),( Yi-xuan Zhang ) 한국미생물 · 생명공학회 2021 Journal of microbiology and biotechnology Vol.31 No.8

The transcriptional capacities of target genes are strongly influenced by promoters, whereas few studies have focused on the development of robust, high-performance and cross-species promoters for wide application in different bacteria. In this work, four novel promoters (P_k.rtufB, P_k.r1, P_k.r2, and P_k.r3) were predicted from Ketogulonicigenium robustum and their inconsistency in the -10 and -35 region nucleotide sequences indicated they were different promoters. Their activities were evaluated by using green fluorescent protein (gfp) as a reporter in different species of bacteria, including K. vulgare SPU B805, Pseudomonas putida KT2440, Paracoccus denitrificans PD1222, Bacillus licheniformis and Raoultella ornithinolytica, due to their importance in metabolic engineering. Our results showed that the four promoters had different activities, with P_k.r1 showing the strongest activity in almost all of the experimental bacteria. By comparison with the commonly used promoters of E. coli (tufB, lac, lacUV5), K. vulgare (Psdh, Psndh) and P. putida KT2440 (JE111411), the four promoters showed significant differences due to only 12.62% nucleotide similarities, and relatively higher ability in regulating target gene expression. Further validation experiments confirmed their ability in initiating the target minCD cassette because of the shape changes under the promoter regulation. The overexpression of sorbose dehydrogenase and cytochrome c551 by P_k.r1 and P_k.r2 resulted in a 22.75% enhancement of 2-KGA yield, indicating their potential for practical application in metabolic engineering. This study demonstrates an example of applying bioinformatics to find new biological components for gene operation and provides four novel promoters with broad suitability, which enriches the usable range of promoters to realize accurate regulation in different genetic backgrounds.

A Wide Dynamic Range NUC Algorithm for IRCS Systems

Li-Hua Cai,Feng-Yun He,Song-Tao Chang,Zhou Li 한국물리학회 2018 THE JOURNAL OF THE KOREAN PHYSICAL SOCIETY Vol.73 No.12

Uniformity is a key feature of state-of-the-art infrared focal planed array (IRFPA) and infrared imaging system. Unlike traditional infrared telescope facility, a ground-based infrared radiant char- acteristics measurement system with an IRFPA not only provides a series of high signal-to-noise ratio (SNR) infrared image but also ensures the validity of radiant measurement data. Normally, a long integration time tends to produce a high SNR infrared image for infrared radiant charac- teristics radiometry system. In view of the variability of and uncertainty in the measured target's energy, the operation of switching the integration time and attenuators usually guarantees the gual- ity of the infrared radiation measurement data obtainted during the infrared radiant characteristics radiometry process. Non-uniformity correction (NUC) coecients in a given integration time are often applied to a specified integration time. If the integration time is switched, the SNR for the infrared imaging will degenerate rapidly. Considering the effect of the SNR for the infrared image and the infrared radiant characteristics radiometry above, we propose a-wide-dynamic-range NUC algorithm. In addition, this essasy derives and establishes the mathematical modal of the algorithm in detail. Then, we conduct verification experiments by using a ground-based MWIR(Mid-wave Infared) radiant characteristics radiometry system with an 400 mm aperture. The experimental results obtained using the proposed algorithm and the traditional algorithm for different integration time are compared. The statistical data shows that the average non-uniformity for the proposed algorithm decreased from 0.77% to 0.21% at 2.5 ms and from 1.33% to 0.26% at 5.5 ms. The testing results demonstrate that the usage of suggested algorithm can improve infrared imaging quality and radiation measurement accuracy.

A New Ferulic Acid Ester and Other Constituents from Dracocephalum peregrinum

Li-Min Dai,Chun-Chao Zhao,Hui-zi Jin,Yun-Heng Shen,Hui-Liang Li,Cai-Yun Peng,Jian Tang,Wei-Dong Zhang 대한약학회 2008 Archives of Pharmacal Research Vol.31 No.10

A new ferulic acid ester, 1'-methyl-2'-hydroxyethyl ferulate (1), together with methylcaffeate (2), 4- hydroxy cinnamic acid (3), ferulic acid (4), caffeic acid (5), diosmetin (6), luteolin (7), 5,3',4'-trihydroxy- 3,7-dimethoxyflavone (8), eriodictyol (9), kaempferol (10), quercetin (11), acacetin-7-Oglcopyranoside (12), 4-(β-glucopyranosyloxy) benzoic acid (13), luteolin-7-O-(6''-feruloyl) glucopyranoside (14), luteolin-7-O-glucopyranoside (15), kaempferide-3-O-rhamnopyranoside (16), quercitrin (17), kaempferol-3-O-glucopyranoside (18), prunasin (19), quercetin-7-O-glucopyranoside (20), quercetin-3-O-glucopyranoside (21), plantaginin (22), linarin (23), luteolin-7-O-rutinoside (24), and chlorogenic acid (25) were isolated from the aerial parts of Dacocephalum peregrinum. The structure of 1 was elucidated on the basis of spectroscopic and HR-ESI-MS analyses. In addition, compound 1 exhibited mild inhibitory effect on NO production in LPS-stimulated RAW264.7 cells.

Combined Expression of Metastasis Related Markers Naa10p, SNCG and PRL-3 and its Prognostic Value in Breast Cancer Patients

Min, Li,Ma, Ruo-Lan,Yuan, Hua,Liu, Cai-Yun,Dong, Bing,Zhang, Cheng,Zeng, Yan,Wang, Li,Guo, Jian-Ping,Qu, Li-Ke,Shou, Cheng-Chao Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.7

Combinations of multiple biomarkers representing distinct aspects of metastasis may have better prognostic value for breast cancer patients, especially those in late stages. In this study, we evaluated the protein levels of N-${\alpha}$-acetyltransferase 10 protein (Naa10p), synuclein-${\gamma}$ (SNCG), and phosphatase of regenerating liver-3 (PRL-3) in 365 patients with breast cancer by immunohistochemistry. Distinct prognostic subgroups of breast cancer were identified by combination of the three biomarkers. The Naa10p+SNCG-PRL-3-subgroup showed best prognosis with a median distant metastasis-free survival (DMFS) of 140 months, while the Naa10p-SNCG+PRL-3+subgroup had the worst prognosis with a median DMFS of 60.5 months. Multivariate analysis indicated Naa10p, SNCG, PRL-3, and the TNM classification were all independent prognostic factors for both DMFS and overall survival (OS). The three biomarker combination of Naa10p, SNCG and PRL-3 performed better in patients with lymph node metastasis, especially those with more advanced tumors than other subgroups. In conclusion, the combined expression profile of Naa10p, SNCG and PRL-3, alone or in combination with the TNM classification system, may provide a precise estimate of prognosis of breast cancer patients.

Comparison of drought resistance of rootstocks 'M9-T337' and 'M26' grafted with 'Huashuo' apple

Shi Cai-Yun,Liu Li,Li Qiu-Li,Wei Zhi-Feng,Gao Deng-Tao 한국원예학회 2022 Horticulture, Environment, and Biotechnology Vol.63 No.3

Drought stress is one of the main limiting factors in apple (Malus domestica Borkh.) cultivation. Rootstock plays an important role in the drought tolerance of apple plants. ‘M.9-T337’ is a novel apple rootstock that was recently introduced and widely cultivated in China. In this study, we selected the new, widely promoted Chinese apple variety ‘Huashuo’ as the scion and grafted it onto ‘M.9-T337’ (HM9). Another combination, ‘Huashuo’/‘M.26’/Malus robusta Rehd. (HM26), served as the experimental control to analyse drought resistance in the two hybrids. We believe that this empirical approach is more representative than merely studying rootstock seedlings. After sustained drought stress for over 1 month, the leaf relative water content had decreased in both types of plants, but to a lesser extent in HM26 than in HM9. The SPAD values increased in both plants, but without significant difference. Drought stress reduced photosynthetic activity in both plants, and the net photosynthetic rate was higher in HM26 than in HM9. The observed changes in the chlorophyll fluorescence parameters indicated that drought had damaged the PSII activity centres of both plants, photosynthetic electron transfer was inhibited, and excessive excitation energy accumulated. However, compared to HM26, HM9 displayed lower maximal PSII quantum photochemical efficiency and potential PSII activity. Moreover, HM9 showed lower antioxidant enzyme activity than HM26 under drought stress. A membership function analysis confirmed that ‘M.9-T337’ was less drought resistant than ‘M.26’. Nevertheless, ‘M.9-T337’ could still recover after prolonged drought stress, indicating it also had good drought resistance.

Oleanolic Acid Inhibits Neuronal Pyroptosis in Ischaemic Stroke by Inhibiting miR-186-5p Expression

Shi-Chang Cai,Xiu-Ping Li,Xing Li,Gen-Yun Tang,Li-Ming Yi,Xiang-Shang Hu 한국뇌신경과학회 2021 Experimental Neurobiology Vol.30 No.6

Ischaemic stroke is a common condition leading to human disability and death. Previous studies have shown that oleanolic acid (OA) ameliorates oxidative injury and cerebral ischaemic damage, and miR-186-5p is verified to be elevated in serum from ischaemic stroke patients. Herein, we investigated whether OA regulates miR-186-5p expression to control neuroglobin (Ngb) levels, thereby inhibiting neuronal pyroptosis in ischaemic stroke. Three concentrations of OA (0.5, 2, or 8 μM) were added to primary hippocampal neurons subjected to oxygen–glucose deprivation/reperfusion (OGD/R), a cell model of ischaemic stroke. We found that OA treatment markedly inhibited pyroptosis. qRT–PCR and western blot revealed that OA suppressed the expression of pyroptosis-associated genes. Furthermore, OA inhibited LDH and proinflammatory cytokine release. In addition, miR-186-5p was downregulated while Ngb was upregulated in OA-treated OGD/R neurons. MiR-186-5p knockdown repressed OGD/R-induced pyroptosis and suppressed LDH and inflammatory cytokine release. In addition, a dual luciferase reporter assay confirmed that miR-186-5p directly targeted Ngb. OA reduced miR-186-5p to regulate Ngb levels, thereby inhibiting pyroptosis in both OGD/R-treated neurons and MCAO mice. In conclusion, OA alleviates pyroptosis in vivo and in vitro by downregulating miR-186-5p and upregulating Ngb expression, which provides a novel theoretical basis illustrating that OA can be considered a drug for ischaemic stroke.

Solving point burnup equations by Magnus method

Yun Cai,Xingjie Peng,Qing Li,Lin Du,Lingfang Yang 한국원자력학회 2019 Nuclear Engineering and Technology Vol.51 No.4

The burnup equation of nuclides is one of the most equations in nuclear reactor physics, which isgenerally coupled with transport calculations. The burnup equation describes the variation of the nuclideswith time. Because of its very stiffness and the need for large time step, this equation is solved byspecial methods, for example transmutation trajectory analysis (TTA) or the matrix exponential methodswhere the matrix exponential is approximated by CRAM. However, TTA or CRAM functions well whenthe flux is constant. In this work, a new method is proposed when the flux changes. It's an improvedmethod compared to TTA or CRAM. Furtherly, this new method is based on TTA or CRAM, and it is moreaccurate than them. The accuracy and efficiency of this method are investigated. Several cases are usedand the results show the accuracy and efficiency of this method are great.

P53 transcription-independent activity mediates selenite-induced acute promyelocytic Leukemia NB4 cell apoptosis

( Li Ying Guan ),( Fang Huang ),( Zhu Shi Li ),( Bing She Han ),( Qian Jiang ),( Yun Ren ),( Yang Yang ),( Cai Min Xu ) 생화학분자생물학회 2008 BMB Reports Vol.41 No.10

Development of a New Thermal Insulating Concrete Block by Spent Polystyrene Foam

Cai, Li Peng,Yun, Ying Wei,Jang, Ii Young,Kim, Seong Kyum Trans Tech Publications, Ltd. 2010 Key Engineering Materials Vol.450 No.-

<P>In this research, a new Thermal Insulating Concrete Block (TICB) is produced via “sandwich” structure by spent polystyrene foam, cement and other raw materials. According to the concerned China Standard Test Method for Building Material, the main properties of this TICB are measured successfully. By comparing its properties with those of autoclaved aerated concrete block as wall material, it can be concluded that comprehensive properties of this concrete block is better than those of autoclaved aerated concrete block. The extrusive advantage of producing this TICB is that large amount of spent polystyrene foam which is hard to be decomposed as industrial waste can be consumed. This is propitious to environmental protection and energy conservation. Therefore, TICB studied in this paper is one kind of green environmental wall material with bright prospect.</P>

Identification of high-affinity VEGFR3-binding peptides through a phage-displayed random peptide library

Li-Feng Shi,Yan Wu,Cai-Yun Li 대한부인종양학회 2015 Journal of Gynecologic Oncology Vol.26 No.4

Objective: Vascular endothelial growth factor (VEGF) interaction with its receptor, VEGFR-3/Flt-4, regulates lymphangiogenesis. VEGFR-3/Flt-4 expression in cancer cells has been correlated with clinical stage, lymph node metastasis, and lymphatic invasion. The objective of this study is to identify a VEGFR-3/Flt-4-interacting peptide that could be used to inhibit VEGFR-3 for ovarian cancer therapy. Methods: The extracellular fragment of recombinant human VEGFR-3/Flt-4 (rhVEGFR-3/Flt-4) fused with coat protein pIII was screened against a phage-displayed random peptide library. Using affinity enrichment and enzyme-linked immunosorbent assay (ELISA) screening, positive clones of phages were amplified. Three phage clones were selected after four rounds of biopanning, and the specific binding of the peptides to rhVEGFR-3 was detected by ELISA and compared with that of VEGF-D. Immunohistochemistry and immunofluorescence analyses of ovarian cancer tissue sections was undertaken to demonstrate the specificity of the peptides. Results: After four rounds of biopanning, ELISA confirmed the specificity of the enriched bound phage clones for rhVEGFR-3. Sequencing and translation identified three different peptides. Non-competitive ELISA revealed that peptides I, II, and III had binding affinities for VEGFR-3 with Kaff (affinity constant) of 16.4±8.6 μg/mL (n=3), 9.2±2.1 μg/mL (n=3), and 174.8±31.1 μg/ mL (n=3), respectively. In ovarian carcinoma tissue sections, peptide III (WHWLPNLRHYAS), which had the greatest binding affinity, also co-localized with VEGFR-3 in endothelial cells lining lymphatic vessels; its labeling of ovarian tumors in vivo was also confirmed. Conclusion: These finding showed that peptide III has high specificity and activity and, therefore, may represent a potential therapeutic approach to target VEGF-VEGFR-3 signaling for the treatment or diagnosis of ovarian cancer.